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However, one disadvantage of such methods is their need to undergo technical revalidation each time a new marker is added to the system. Moreover, polymorphic markers cannot be assessed at the single-nucleotide level in these assays. Since new<jats:named-content content-type=\"genus-species\">Brucella<\/jats:named-content>species are continuously being described, open methodologies able to accommodate new markers while preserving all other system parameters have an obvious advantage. We present a ligase chain reaction (LCR)-based method that simultaneously assesses multiple genetic markers at the single-nucleotide level. Most of the selected markers originate from a multilocus sequence typing (MLST) database that has been extensively validated on hundreds of different<jats:named-content content-type=\"genus-species\">Brucella<\/jats:named-content>strains. When assayed on both reference and field strains, the method yields characteristic capillary electrophoresis profiles for each of the 10<jats:named-content content-type=\"genus-species\">Brucella<\/jats:named-content>species described to date and displays discriminatory potential below the species level for some. Since the LCR methodology is insensitive to interference resulting from the use of multiple oligonucleotides in a single mixture, the way is open for smooth future updates of the proposed system. Such updates are inevitable, given the pending description of new<jats:named-content content-type=\"genus-species\">Brucella<\/jats:named-content>species.<\/jats:p>","DOI":"10.1128\/aem.00767-11","type":"journal-article","created":{"date-parts":[[2011,7,30]],"date-time":"2011-07-30T04:16:14Z","timestamp":1311999374000},"page":"6674-6679","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":12,"title":["Nucleotide Polymorphism-Based Single-Tube Test for Robust Molecular Identification of All Currently Described Brucella Species"],"prefix":"10.1128","volume":"77","author":[{"given":"Pierre","family":"Wattiau","sequence":"first","affiliation":[{"name":"Veterinary and Agrochemical Research Centre, Department of Bacterial Diseases, Groeselenberg 99, B-1180 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian M.","family":"Whatmore","sequence":"additional","affiliation":[{"name":"Veterinary Laboratories Agency, Department of Bacteriology, Addlestone, Surrey KT15 3NB, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mieke","family":"Van Hessche","sequence":"additional","affiliation":[{"name":"Veterinary and Agrochemical Research Centre, Department of Bacterial Diseases, Groeselenberg 99, B-1180 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacques","family":"Godfroid","sequence":"additional","affiliation":[{"name":"Norwegian School of Veterinary Science, Department of Food Safety and Infection Biology, Section of Arctic Veterinary Medicine, Stakkevollveien 23, 9016 Troms\u00f8, Norway"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Fretin","sequence":"additional","affiliation":[{"name":"Veterinary and Agrochemical Research Centre, Department of Bacterial Diseases, Groeselenberg 99, B-1180 Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1515\/CCLM.2007.305"},{"key":"e_1_3_3_3_2","unstructured":"AltonG. 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